Fluorescence polarization as an analytical tool to evaluate galectin-ligand interactions.
(2004) In Analytical Biochemistry 334(1). p.36-47- Abstract
- Galectins are a family of β-galactose binding lectins associated with functions such as immunological and malignant events. To study the binding affinity of galectins for natural and artificial saccharides and glycoconjugates we have developed an assay using fluorescence polarization. A collection of fluorescein-conjugated saccharides was synthesized and used as probes with galectins-1 and -3 and the two carbohydrate recognition domains of galectin-4. Direct binding of a fixed probe amount with different amounts of each galectin defined specificity and selectivity and permitted selection of the optimal probe for inhibition studies. Then fixed amounts of galectin and selected probe were used to screen the inhibitory potency of a library of... (More)
- Galectins are a family of β-galactose binding lectins associated with functions such as immunological and malignant events. To study the binding affinity of galectins for natural and artificial saccharides and glycoconjugates we have developed an assay using fluorescence polarization. A collection of fluorescein-conjugated saccharides was synthesized and used as probes with galectins-1 and -3 and the two carbohydrate recognition domains of galectin-4. Direct binding of a fixed probe amount with different amounts of each galectin defined specificity and selectivity and permitted selection of the optimal probe for inhibition studies. Then fixed amounts of galectin and selected probe were used to screen the inhibitory potency of a library of nonfluorescent compounds. As the assay is in solution and does not require separation of free and bound probe, it is simple and rapid and can easily be applied to different unlabeled galectins. As all interaction components are known, Kd values for galectin–inhibitor interaction can be directly calculated without approximation other than the assumption of a simple one-site competition. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/129925
- author
- Sörme, Pernilla LU ; Kahl-Knutsson, Barbro ; Huflejt, Margaret ; Nilsson, Ulf LU and Leffler, Hakon LU
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Analytical Biochemistry
- volume
- 334
- issue
- 1
- pages
- 36 - 47
- publisher
- Elsevier
- external identifiers
-
- wos:000224710600004
- pmid:15464951
- scopus:4644260095
- pmid:15464951
- ISSN
- 1096-0309
- DOI
- 10.1016/j.ab.2004.06.042
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Organic chemistry (S/LTH) (011001240), Division of Microbiology, Immunology and Glycobiology - MIG (013025200)
- id
- b61bce62-d583-449d-bab6-2fa27d301ed0 (old id 129925)
- alternative location
- http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15464951&dopt=Abstract
- date added to LUP
- 2016-04-01 11:34:52
- date last changed
- 2022-04-12 22:13:06
@article{b61bce62-d583-449d-bab6-2fa27d301ed0,
abstract = {{Galectins are a family of β-galactose binding lectins associated with functions such as immunological and malignant events. To study the binding affinity of galectins for natural and artificial saccharides and glycoconjugates we have developed an assay using fluorescence polarization. A collection of fluorescein-conjugated saccharides was synthesized and used as probes with galectins-1 and -3 and the two carbohydrate recognition domains of galectin-4. Direct binding of a fixed probe amount with different amounts of each galectin defined specificity and selectivity and permitted selection of the optimal probe for inhibition studies. Then fixed amounts of galectin and selected probe were used to screen the inhibitory potency of a library of nonfluorescent compounds. As the assay is in solution and does not require separation of free and bound probe, it is simple and rapid and can easily be applied to different unlabeled galectins. As all interaction components are known, Kd values for galectin–inhibitor interaction can be directly calculated without approximation other than the assumption of a simple one-site competition.}},
author = {{Sörme, Pernilla and Kahl-Knutsson, Barbro and Huflejt, Margaret and Nilsson, Ulf and Leffler, Hakon}},
issn = {{1096-0309}},
language = {{eng}},
number = {{1}},
pages = {{36--47}},
publisher = {{Elsevier}},
series = {{Analytical Biochemistry}},
title = {{Fluorescence polarization as an analytical tool to evaluate galectin-ligand interactions.}},
url = {{http://dx.doi.org/10.1016/j.ab.2004.06.042}},
doi = {{10.1016/j.ab.2004.06.042}},
volume = {{334}},
year = {{2004}},
}